This invention has identified novel peptide epitopes recognized by the B cell lymphoma-reactive Lym-1 antibody. Because Lym-1 specifically reacts with non-Hodgkin""s B cell lymphoma cells, the invention provides an improved, accurate means to identify cancer patients potentially responsive to Lym-1 antibody used as a cytotoxic therapeutic reagent. The invention also provides methods of generating antibodies directed against non-Hodgkin""s B cell lymphoma cells which can be used in the treatment of non-Hodgkin""s B cell lymphoma.
Low grade B-cell non-Hodgkin""s lymphomas (B-NHL) represent a markedly heterogeneous group of lymphoproliferative disorders (Gaidano (1997) Leuk. Lymphoma 26 Suppl. 1:107-113; Gandini (1996) Cancer Genet Cytogenet. 86:120-123). A widely used treatment for these lymphomas involves administration of a B-NHL-specific antibody, called Lym-1. Lym-1 is a murine IgG2a monoclonal antibody. When conjugated to cytotoxic agents, Lym-1 targets and kills B-NHL lymphoma cells (see, e.g., Rose (1996) Cancer Immunol. Immunother. 43:26-30; Epstein (1987) Cancer Res. 47:830-840). Lym-1 has been radiolabeled with 131I (see, e.g., DeNardo (1997) Cancer 80:2706-2711) and conjugated to the ribosome inactivating protein gelonin (see, e.g., O""Boyle (1995) J. Immunother. Emphasis Tumor Immunol. 18:221-230). However, these reagents are inherently toxic, and not all B-NHL patients have Lym-1 reactive cancer cells. Thus, there is a need for a means to identify which patients will be responsive to such immunotherapy. Wile Lym-1 binding is associated with the expression of HLA DR10 by a patient, unfortunately, the absence of DR10, does not consistently correlate with the presence or absence of the Lym-1 reactive epitope. In another words, Lym-1 can react with HLA DR molecules other than DR10. Thus, there is a great need for a means to quickly, efficiently, and accurately determine the presence of a Lym-1 reactive epitope in a B-NHL cancer patient. The present invention, which for the first time identifies Lym-1 reactive peptide epitopes, fulfills these and other needs.
Typically, no immune response is generated by the cancer patient against B-NHL cells. However, based on studies with other tumor specific antigens (e.g., PSA antigen in prostate cancer), identification of an immunogenic peptide, followed by its administration with adjuvant, can elicit a tumor-specific immune response (see, e.g., Correale (1998) J. Immunol. 161:3186-3194). See also, Gjertsen (1998) Vox Sang. 74 Suppl 2:489-495, who uses an immunogenic peptide from a carcinogenic, mutant ras polypeptide to generate an immune response to pancreatic- and colorectal adenocarcinomas. The present invention, by identifying Lym-1 reactive epitopes on B-NHL cells, provides such a therapeutic immunogenic peptide.
The invention for the first time provides a composition comprising an isolated or recombinant peptide comprising a subsequence of a Class II major histocompatibility molecule that generates an immune response to a non-Hodgkin""s B cell lymphoma cell. The peptide of the invention has a structure comprising R1-R2-R3-R4-R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16, wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 and R4 are members independently selected from the group consisting of all amino acids; R5 is Ala, Glu, Asp, Val, Leu or Ile; R6 and R7 are members independently selected from the group consisting of all amino acids; R8 is Thr; R9, R10, R11, R12, R13, R14, and R15 are members independently selected from the group consisting of all amino acids; and, R16 is Val.
In one embodiment, the peptide of the invention has a structure wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is selected from the group consisting of all amino acids; R5 is Ala; R6 and R7 are members independently selected from the group consisting of all amino acids; R8 is Thr; R9 is selected from the group consisting of all amino acids; R10 is Cys; R11, R12, R13, R14, and R15 are members independently selected from the group consisting of all amino acids; and, R16 is Val (SEQ ID NO:1). In a preferred embodiment, the immunogenic peptide comprises a structure wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R, is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2).
In alternative embodiments, the composition further comprises a pharmaceutically acceptable excipient and an adjuvant.
The composition of the invention can generate an immune response to a non-Hodgkin""s lymphoma cell (B-NHL), including, e.g., a B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CCL/SLL) cell, a lymphoplasmacytoid lymphoma (LPL) cell, a follicular lymphoma (FL) cell, a mucosa-associated lymphoid tissue lymphoma (MALTL) cell, a splenic lymphoma with villous lymphocytes (SLVL) cell and a mantle cell lymphoma cell.
The invention also provides a method for detecting a nucleic acid in a biological sample, wherein the nucleic acid encodes a peptide capable of specifically binding to a Lym-1 antibody. The method of the invention comprises contacting the sample with an oligonucleotide primer pair capable of amplifying a subsequence of an MHC nucleic acid, which subsequence encodes a polypeptide comprising a peptide of the invention, as described above; amplifying the nucleic acid; and, detecting the amplified nucleic acid. In alternative embodiments, the MHC gene is HLA-DR 10 and the subsequence encodes a peptide wherein R1 is Gin, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2).
In the methods, the biological sample can comprise a B cell, or specifically, a B lymphocytic non-Hodgkin""s lymphoma cell (B-NHL). The B-NHL cell can be a B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CCL/SLL) cell, a lymphoplasmacytoid lymphoma (LPL) cell, a follicular lymphoma (FL) cell, a mucosa-associated lymphoid tissue lymphoma (MALTL) cell, a splenic lymphoma with villous lymphocytes (SLVL) cell and a mantle cell lymphoma cell. In alternative embodiments the biological sample can be a body fluid sample or a biopsy sample; and the body fluid sample can be a blood sample.
The invention further provides a kit for detecting a nucleic acid in a biological sample, wherein the nucleic acid encodes a peptide capable of specifically binding to a Lym-1 antibody. The kit comprises an oligonucleotide primer pair capable of amplifying a subsequence of an MHC gene or gene product, which subsequence encodes a polypeptide comprising a peptide of the invention. In alternative embodiments, the MHC gene can be HLA-DR 10; and, the peptide can comprise a structure wherein R1 is Gin, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2).
In another embodiment, the kit of the invention can further comprise an instructional material teaching a use of the kit, wherein the instructional material indicates that the kit is used for the detection of nucleic acid encoding a peptide reactive with a Lym-1 antibody and that the polypeptide is associated with non-Hodgkin""s B cell lymphomas.
The invention also provides a method for detecting an antibody reactive with a non-Hodgkin""s B cell lymphoma (B-NHL) cell. The method comprises contacting a sample, which can be a biological sample, with a composition of the invention under immunologically reactive conditions, and then detecting whether an antibody has specifically bound to the composition. In one embodiment, the composition comprises a peptide having a structure wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2). In various embodiments of this method, the antibody is generated by a recombinant nucleic acid library, the recombinant nucleic acid is a phage display library, and the composition is fixed to a solid surface.
The invention further provides a method for generating an antibody reactive with a non-Hodgkin""s B cell lymphoma (B-NHL) cell. The method comprises administering an immunogenically effective amount of a composition of the invention to a mammal. The composition can comprise a peptide having a structure wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Gys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2). The B-NHL cell can be a B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CCL/SLL) cell, a lymphoplasmacytoid lymphoma (LPL) cell, a follicular lymphoma (FL) cell, a mucosa-associated lymphoid tissue lymphoma (MALTL) cell, a splenic lymphoma with villous lymphocytes (SLVL) cell and a mantle cell lymphoma cell.
The invention provides an immunogenic composition capable of eliciting an immunogenic response directed to a polypeptide epitope, wherein the epitope comprises an amino acid sequence having a structure comprising R1-R2-R3-R4-R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16, wherein R1 is Gln, Lys, or Arg; R2 is Arg, R3 and R4 are members independently selected from the group consisting of all amino acids; R5 is Ala, Glu, Asp, Val, Leu or Ile; R6 and R7 are members independently selected from the group consisting of all amino acids; R8 is Thr; R9, R10, R11, R12, R13, R14, and R15 are members independently selected from the group consisting of all amino acids; and, R16 is Val. In one embodiment, the epitope comprises a sequence wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2). The immunogenic response can generate antibodies (i.e., a humoral response) specific for the polypeptide epitope. Alternatively, the immunogenic response can generate an epitope specific cellular response.
The invention further provides a method of inducing an immunogenic response directed to a polypeptide epitope, comprising administering an immunogenically effective amount of a composition comprising a polypeptide epitope to a mammal, wherein the epitope comprises an amino acid sequence having a structure comprising R1-R2-R3-R4-R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16, wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 and R4 are members independently selected from the group consisting of all amino acids; R5 is Ala, Glu, Asp, Val, Leu or Ile; R6 and R7 are members independently selected from the group consisting of all amino acids; R8 is Thr; R9, R10, R11, R12, R13, R14, and R15 are members independently selected from the group consisting of all amino acids; and, R16 is Val. In one embodiment, the epitope comprises an amino acid sequence having a structure wherein R1 is Gln, Lys, or Arg; R2 is Arg; R3 is Arg; R4 is Ala; R5 is Ala; R6 is Val; R7 is Asp; R8 is Thr; R9 is Tyr; R10 is Cys; R11 is Arg; R12 is His; R13 is Asn; R14 is Tyr; R15 is Gly, and R16 is Val (SEQ ID NO:2). The immunogenic response can generate antibodies (i.e., a humoral response) specific for the polypeptide epitope. Alternatively, the immunogenic response can generate an epitope specific cellular response. In various embodiments, the method involves administering the immunogenic composition to a human, a mouse or a rabbit.
A further understanding of the nature and advantages of the present invention may be realized by reference to the remaining portions of the specification, the figures and claims.
All publications, patents and patent applications cited herein are hereby expressly incorporated by reference for all purposes.